JP4584919B2 - Analytical specimen - Google Patents

Description

  According to the present invention, when a sample containing a substance to be analyzed is introduced, the sample comes into contact with and reacts with a detection component contained in a reaction portion to generate a detectable substance (signal substance) or a detectable characteristic ( The present invention relates to an analytical test piece capable of presenting signal characteristics. More specifically, it is useful as an inspection chip for inspecting and analyzing the properties of samples containing substances to be analyzed (for example, body fluids of humans and animals, especially urine, blood, etc.). In addition, the present invention relates to a high quality (high accuracy, high density, high sensitivity, etc.) test specimen for analysis.

As an analytical test piece for examining and analyzing the properties of a sample containing a substance to be analyzed (for example, body fluids of humans and animals, particularly urine, blood, etc.), for example, a porous structure having good absorbability ( A porous membrane and a method for producing the same are disclosed in which a sample is uniformly absorbed in a test portion comprising a porous layer, a porous membrane, etc., and a liquid junction with an adjacent test portion is prevented (see Patent Document 1). . Further, an analytical test piece is disclosed in which one or more test parts having a detection part for detecting a detectable substance are provided, and the detection part contains a layered inorganic compound (such as synthetic smectite) (Patent Document 2). reference).
Japanese Patent Laid-Open No. 2-6541 Japanese Patent Laid-Open No. 9-184837

  However, in the porous membrane, the analytical test piece and the production method thereof disclosed in Patent Document 1 and Patent Document 2, a plurality of types of detection components are impregnated in a single buffer in the reaction portion. In addition to the rapid deterioration of detection components, lack of stability, low reaction efficiency, and problems in analysis reliability, high precision, high density, high sensitivity, etc., which have recently been required for test specimens for analysis, etc. However, it was not always satisfactory.

  The present invention has been made in view of the above-described problems, and is used as a test chip for inspecting and analyzing properties of a sample containing a substance to be analyzed (for example, body fluids of humans and animals, particularly urine, blood, etc.). An object of the present invention is to provide a test specimen for analysis that is useful and capable of performing a precise inspection with a small amount of sample and that has high quality (high accuracy, high density, high sensitivity, etc.).

  In order to achieve the above object, according to the present invention, the following analytical test strip is provided.

[1] A carrier and a reaction portion containing a detection component disposed on and / or in the surface of the carrier, and a sample containing the substance to be analyzed is introduced onto the surface of the carrier. When contact with the detecting components contained in the reaction section, react to a analytical test strip capable of generating or detectable characteristics of presentation of detectable objects substance, said in said reaction section the content of the detection component, in from one end of the reaction section to the other, which becomes configured to increase or decrease continuously, the detection component in the reaction portion, as a set of spots together becomes disposed in a predetermined arrangement pattern, the arrangement density of the spots constituting the arrangement pattern is made the in from one end of the reaction portion to the other end, it is configured to increase or decrease continuously analyzed for Kenhen (hereinafter sometimes referred to as "first invention").

[2] The detection component in the reaction portion is composed of a plurality of types that are spaced apart from each other and are independently arranged, and the detection component is contained in the detection component from one end to the other end of the reaction portion. The test specimen for analysis according to [1], which is configured to continuously increase or decrease for each type.

[3] The content of the detected components of spots constituting the arrangement pattern, in from one end of the reaction section to the other, consisting configured to increase or decrease continuously the [1] or [ 2] The test specimen for analysis as described in 2].

[4] the size of the spots constituting the arrangement pattern, in from one end of the reaction portion to the other end, one of the composed configured to increase or decrease continuously [1] to [3] The analytical test piece according to Crab.

[5] in the detection component of spots constituting the arrangement pattern, the content in the thickness direction of the carrier, in from one end of the reaction portion to the other end, is configured to increase or decrease continuously The test specimen for analysis according to any one of the above [1] to [4].

[6] A carrier and a reaction portion containing a detection component disposed on and / or in the surface of the carrier, and a sample containing the substance to be analyzed is introduced onto the surface of the carrier When contact with the detecting components contained in the reaction section, react to a analytical test strip capable of generating or detectable characteristics of presentation of detectable objects substance, said reaction portion has multiple And the content of the detection component in the reaction portion is stepwise adjacent to each other for each of the plurality of reaction sites from one end to the other end of the reaction portion, or spaced apart from each other, and independently fragmentary, which become configured to increase or decrease, the detection component in the reaction portion, it becomes disposed in a predetermined arrangement pattern as a set of spots , The array pattern Arrangement density of spots constituting the emission is at one end of the reaction portion to the other end, for each of the plurality of the reaction sites that divide the reaction part, fragmentarily stepwise or discontinuously in succession, Analytical test strip configured to increase or decrease (hereinafter sometimes referred to as “second invention”).

[ 7 ] The detection component in the reaction portion is composed of a plurality of types that are spaced apart from each other and are independently arranged, and the detection component is contained in the detection component from one end to the other end of the reaction portion. [ 6 ], wherein each of the plurality of reaction sites corresponding to each of the plurality of types is configured to increase or decrease stepwise adjacent to each other, or separated from each other, independently, fragmentarily. The test specimen for analysis as described.

[8] The content of the detected components of spots constituting the arrangement pattern, in from one end of the reaction portion to the other end, for each of the plurality of the reaction sites that divide the reaction portion, stepwise continuously The analytical test strip according to [6] or [7], wherein the test strip is configured to increase or decrease in a continuous or discontinuous manner.

[9] the size of the arrangement of the spots constituting the arrangement pattern, in from one end of the reaction portion to the other end, for each of the plurality of the reaction sites that divide the reaction portion, stepwise or continuously The analytical test strip according to any one of [6] to [8], which is configured to discontinuously and fragmentally increase or decrease.

[10] in the detection component of spots constituting the arrangement pattern, the content in the thickness direction of the carrier, in from one end of the reaction portion to the other end, a plurality of the reaction sites that divide the reaction portion The analytical test strip according to any one of [6] to [9], wherein the analytical test strip is configured to increase or decrease continuously, stepwise, discontinuously, or piecewise.

[11] wherein said carrier, said a fibrous body or a porous body [1] The analytical test strip of any one of - [10].

[ 12 ] The method according to any one of [1] to [ 11 ], wherein the reaction portion containing the detection component is disposed on and / or inside the surface of the carrier using an inkjet method. Test specimen for analysis.

  According to the present invention, it is useful as a test chip for inspecting and analyzing the properties of samples containing substances to be analyzed (for example, body fluids of humans and animals, especially urine, blood, etc.), and a precise test is performed with a small amount of samples. In addition, an analytical test strip of high quality (high accuracy, high density, high sensitivity, etc.) can be provided.

It is explanatory drawing which shows typically the basic example (1st basic example) of the test piece for analysis of 1st invention. It is explanatory drawing which shows typically the one modification (1st modification 1) which changed the reaction part in the test piece for analysis (1st basic example) of 1st invention shown in FIG. FIG. 1 schematically shows an enlarged part (A part) of another modified example (first modified example 2) of the reaction portion in the analytical test piece (first basic example) of the first invention shown in FIG. FIG. FIG. 1 schematically shows an enlarged part (part A) of another modified example (first modified example 3) of the reaction part in the analytical test piece (first basic example) of the first invention shown in FIG. FIG. FIG. 1 schematically shows an enlarged part (part A) of another modified example (first modified example 4) of the reaction part in the analytical test piece (first basic example) of the first invention shown in FIG. FIG. FIG. 1 schematically shows an enlarged part (part A) of another modified example (first modified example 5) of the reaction part in the analytical test piece (first basic example) of the first invention shown in FIG. FIG. FIG. 1 schematically shows an enlarged part (part A) of another modified example (first modified example 6) of the reaction part in the analytical test piece (first basic example) of the first invention shown in FIG. FIG. It is sectional drawing of the thickness direction of the support | carrier in the AA line | wire of Fig.7 (a). It is explanatory drawing which shows typically one basic example (2nd basic example 1) of the test piece for analysis of 2nd invention. FIG. 9 is a partially enlarged view of a reaction portion (B portion) in the analytical test piece (second basic example 1) of the second invention shown in FIG. It is explanatory drawing which shows typically the other basic example (2nd basic example 2) of the test piece for analysis of 2nd invention. FIG. 10 is a partially enlarged view of a reaction portion (D portion) in the analytical test piece (second basic example 2) of the second invention shown in FIG. 9 (a). It is explanatory drawing which shows typically the one modification (2nd modification 1) which changed the reaction part in the test piece for analysis (2nd basic example 1) of 2nd invention shown to Fig.8 (a). FIG. 8A is a schematic enlarged view of a part (C portion) of another modified example (second modified example 2) of the reaction part in the analytical test piece (second basic example 1) of the second invention shown in FIG. FIG. Partial enlargement schematically showing another modification (second modification 3) of the reaction part (B part) in the analytical test piece (second basic example 1) of the second invention shown in FIG. 8 (a). FIG. Partial enlargement schematically showing another modification (second modification 4) of the reaction part (B part) in the analytical test piece (second basic example 1) of the second invention shown in FIG. 8 (a). FIG. Partial enlargement schematically showing another modification (second modification 5) of the reaction part (B part) in the analytical test piece (second basic example 1) of the second invention shown in FIG. 8 (a). FIG. Partial enlargement schematically showing another modified example (first modified example 6) of the reaction part (B part) in the analytical test piece (second basic example 1) of the second invention shown in FIG. 8 (a). FIG. It is sectional drawing of the thickness direction of the support | carrier in CC line of Fig.15 (a).

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 ... Carrier, 1a ... Separation part, 2 ... Reaction part, 3 ... Spot (dot), 3a-3m, 3o-3s ... Spot (dot), 4 ... Reaction site, 4a-4s ... Reaction site, 5 ... Reaction site 5a to 5d ... reaction site, 10, 10a, 20, 20a ... test specimen for analysis, X (X (A), X (B)), R (R (A), R (B)) ... of the reaction part One end, Y (Y (A), Y (B)), S (S (A), S (B)) ... the other end of the reaction part.

  Hereinafter, the best mode for carrying out the present invention will be specifically described with reference to the drawings.

FIG. 1 is an explanatory view schematically showing a basic example (first basic example) of an analytical test piece of the first invention. As shown in FIG. 1, the analytical test strip 10 of the first invention includes a carrier 1 and a reaction portion 2 containing a detection component disposed on and / or in the surface of the carrier 1. When a sample (not shown) containing the substance to be analyzed is introduced to the surface of the carrier 1, it comes into contact with and reacts with the detection component contained in the reaction portion 2, and a detectable substance (signal substance) An analytical test piece capable of presenting a detectable or detectable characteristic (signal characteristic), wherein the content of the detected component in the reaction part 2 is from one end (X) to the other end (Y) of the reaction part 2 The detection component in the reaction portion 2 is arranged in a predetermined arrangement pattern (spot pitch) as an aggregate of spots (dots) 3, and is configured to continuously increase or decrease , Spots that make up the array pattern ) Arrangement density of 3, in the end of the reaction section 2 (X) to the other end (Y), and is configured to increase or decrease continuously. FIG. 1 shows a case where the reaction portion 2 is configured to continuously increase from one end (X) to the other end (Y).

  Thus, by providing a difference in the amount per unit area (volume) of the detection component in the reaction portion 2 in the analysis test piece 10, the concentration of the sample containing the analysis target substance can be adjusted with one analysis test piece. It can be detected in analog form, and the sample concentration can be digitized. Therefore, a precise value can be output as a test result with a small amount of sample for one analytical test piece.

  The carrier 1 used in the first invention is not particularly limited as long as it can form a reaction portion by arranging and holding the detection component on and / or inside the surface, but a flat surface Those having the following are preferred. For example, a fibrous body or a porous body can be mentioned as a suitable example. Among these, a hydrophilic fibrous body or a porous body is preferable. As the material, for example, celluloses, polyether sulfones, acrylic polymers and the like are suitable, and the pore diameter is preferably 0.2 μm to several μm. In this way, by using a fibrous body or porous body as the carrier 1, the amount of the detection component forming the reaction portion penetrates into the inside of the carrier 1 and a large number of detection components are retained in the thickness direction. It is possible to obtain a highly sensitive test strip with improved analytical sensitivity.

  The detection component used in the first invention may be any component capable of generating a signal substance or presenting signal characteristics through contact reaction with the analysis target substance in the sample by introducing the sample containing the analysis target substance. There is no restriction | limiting in particular, One type may be individual and multiple types of combinations may be sufficient. In the case of one type alone, for example, when the activity of lactate dehydrogenase is measured as a substance to be analyzed (when a solution containing lactate dehydrogenase is used as a sample), the detection component solution may be lactic acid or complement as a substrate. A solution containing NAD (nicotinamide adenine dinucleotide) as an enzyme, 1-methoxy PMS (phenazine methosulfate) as an electron carrier, and NTB (nitrotetrazolium blue) as a tetrazolium reagent, a phosphate buffer, Tris-HCl (Tris-HCl) What adjusted pH to the predetermined value with buffer solutions, such as a buffer solution, can be mentioned. Further, a predetermined amount of a surfactant may be added to the above solution. By adding the surfactant, the detection component on the surface / inside of the carrier can be uniformly dispersed. Such a surface active agent may be any one of anionic, cationic, nonionic, etc., and can be appropriately selected according to use conditions. As anionic, alkyl allyl sulfonate, alkyl benzene sulfonate, etc., as cationic, alkyl, trimethyl ammonium, alkyl, pyridium, etc., as nonionic, as polyoxyethylene fatty acid ester, polyoxyethylene alkyl phenol Can be cited as a suitable example.

  In the first invention, it is preferable to include a color former in the detection component. When a plurality of types of detection components are used in combination, it is preferable that at least one of them includes a color former. By including a color former in the detection component, it is possible to generate a clear signal substance or to exhibit signal characteristics. Examples of the solution containing such a color former include a tetrazolium reagent as a reducing system and 4-aminoantipyrine, a phenol-based solution as an oxidizing system. Further, when a plurality of types of detection components are used in combination, it is preferable to contain color formers that exhibit different colors for each type. With this configuration, it is possible to easily and accurately determine the inspection result in the visual inspection.

  FIG. 2 is an explanatory view schematically showing one modified example (first modified example 1) in which the reaction portion in the analytical test piece (first basic example) of the first invention shown in FIG. 1 is changed. As shown in FIG. 2, in the first modified example 1, the detection components in the reaction portion 2 of the analytical test strip 10a are composed of a plurality of types that are separated from each other and arranged independently, and contain detection components. The ratio continuously increases or decreases for each type of detection component from one end (X (A) and X (B)) to the other end (Y (A) and Y (B)) of the reaction portion 2. It is configured as follows. FIG. 2 shows a case where two types (A type and B type) of detection components that are spaced apart from each other and independently arranged are used. In FIG. 2, the detection component of type A (same for type B) has a higher content of detection component on the other end (Y (A) and Y (B)) side (lower side in the drawing). It is shown that. Further, FIG. 2 shows a case where two types (A type and B type) of detection components are separated from each other and arranged independently by the separation unit 1a.

  With this configuration, a plurality of inspection items can be accurately inspected with a small amount of sample.

  As an example of such a combination of two types (A type and B type) of detection components that are spaced apart from each other and independently arranged, for example, glucose is measured as the first type (A type) as the analysis target substance. And the second type (B type) that measure cholesterol.

  Specific examples of the above combinations include glucose dehydrogenase, NAD (nicotinamide adenine dinucleotide) as a coenzyme, diaphorase as an electron carrier, MTT (3- (4 , 5-dimethylthiazol-2-yl) -2,5-diphenyl-2H-tetrazolium bromide), and as a solution of the B-type detection component, cholesterol dehydrogenase and NAD as a coenzyme ( Nicotinamide adenine dinucleotide), a solution containing 1-methoxy PMS (phenazine methosulfate) as an electron carrier, NTB (nitrotetrazolium blue) as a tetrazolium reagent, and a predetermined solution with Tris-HCl (Tris-HCl) buffer or the like. List the value adjusted pH It can be. Further, a predetermined amount of a surfactant may be added to the above solution. By adding the surfactant, the detection component on the surface / inside of the carrier can be uniformly dispersed. Such a surface active agent may be any one of anionic, cationic, nonionic, etc., and can be appropriately selected according to use conditions. As anionic, alkyl allyl sulfonate, alkyl benzene sulfonate, etc., as cationic, alkyl, trimethyl ammonium, alkyl, pyridium, etc., as nonionic, as polyoxyethylene fatty acid ester, polyoxyethylene alkyl phenol Can be cited as a suitable example. With the configuration described above, the A-type detection component develops a blue color when glucose is detected, and the B-type detection component develops a red-purple color when cholesterol is detected.

  FIG. 3 is an enlarged schematic view of a part (part A) of another modified example (first modified example 2) of the reaction part in the analytical test piece (first basic example) of the first invention shown in FIG. FIG. As shown in FIG. 3, in the first modification 2, the detection components in the reaction portion 2 are arranged in a predetermined arrangement pattern (spot pitch) as an aggregate of spots (dots) 3. . 3, the other end (Y) side (lower side in the drawing) shown in FIG. 1 indicates that the content ratio of the detected component of each spot (dot) 3 is larger. In this case, the shape of the spot (dot) 3 is not particularly limited. For example, the spot (dot) 3 has a predetermined plane parallel to the surface of the carrier 1 (for example, a plane passing through a portion having a depth of 1/2 from the surface of the carrier 1). Examples of the cut cross-sectional shape include a circular shape, an elliptical shape, an oval shape, and a racing track shape.

  By comprising in this way, a sample can be easily supplied to the detection component arrange | positioned on the support | carrier surface or the inside from the periphery, and it becomes possible to make the arrange | positioned detection component react efficiently. Therefore, sufficient detection sensitivity can be obtained with a small amount of detection component. Further, since the reaction can proceed from the entire surface of the dot, the detection time can be shortened.

  A method of forming spots (dots) and array patterns will be described later.

  FIG. 4 is an enlarged schematic view of a part (part A) of another modified example (first modified example 3) of the reaction part in the analytical test piece (first basic example) of the first invention shown in FIG. FIG. As shown in FIG. 4, in the first modification 3, the content of the detected component of the spot (dot) 3 constituting the arrangement pattern in the reaction portion 2 is changed from one end (X) of the reaction portion 2 to the other end (X). Up to Y), it is configured to continuously increase or decrease. FIG. 4 shows three types of spots (dots) 3a, 3b, and 3c having different detection component contents among the spots (dots) 3, and the content of the spot (dot) 3a is the smallest. The case where the content of (dot) 3c is the largest is shown.

  With this configuration, it is possible to give a change in sensitivity without changing the dot size. Therefore, it can be set as the test piece for analysis of a high dynamic range in a small area.

FIG. 5 is a schematic enlarged view of a part (part A) of another modified example (first modified example 4) of the reaction part in the analytical test piece (first basic example) of the first invention shown in FIG. FIG. As shown in FIG. 5, in the first modification example 4, the arrangement density of spots (dots) 3 constituting the arrangement pattern in the reaction portion 2 is from one end (X) to the other end (Y) of the reaction portion 2. (See FIG. 1), it is configured to continuously increase or decrease. In FIG. 5, four types of spots (dots) 3d, 3e, 3f, and 3g having different arrangement densities are shown among the spots (dots) 3, and the pitches p1, p2, and p3 of the arrangement intervals are the other ends. Although the case where it is denser (p1> p2> p3) on the (Y) side is shown, the pitch of the arrangement interval is varied in the direction perpendicular to the direction from one end (X) to the other end (Y). Alternatively, the pitch of the arrangement interval may be different in both the direction from one end (X) to the other end (Y) and the direction perpendicular thereto.

  FIG. 6 is a schematic enlarged view of a part (part A) of another modified example (first modified example 5) of the reaction part in the analytical test piece (first basic example) of the first invention shown in FIG. FIG. As shown in FIG. 6, in the first modification 5, the size of the spot (dot) 3 constituting the arrangement pattern in the reaction portion 2 is from one end (X) to the other end (Y) of the reaction portion 2. (See FIG. 1), it is configured to continuously increase or decrease. In FIG. 6, three types of spots (dots) 3 h, 3 i, and 3 j having different sizes among the spots (dots) 3 are shown, and each size is larger on the other end (Y) side. (Spot (dot) 3h is the smallest and spot (dot) 3j is the largest).

  By configuring as in the first modified example 4 and the first modified example described above, it is possible to easily make a difference in sensitivity with the same concentration and the same amount of detection component (solution), so that an analytical test with high productivity. While obtaining a piece, it is possible to reliably make a difference in sensitivity within the same test piece.

  FIG. 7A is an enlarged view of a part (part A) of another modified example (first modified example 6) of the reaction part in the analytical test piece (first basic example) of the first invention shown in FIG. FIG. 7 (b) is a cross-sectional view in the thickness direction of the carrier taken along line AA of FIG. 7 (a). As shown in FIG. 7 (a), in the first modified example 6, the content in the thickness direction of the carrier 1 in the detected components of the spots (dots) 3 constituting the arrangement pattern in the reaction portion 2 is the reaction. From one end (X) to the other end (Y) of the portion 2 (see FIG. 1), the portion 2 is configured to increase or decrease continuously. 7A shows three types of spots (dots) 3k, 3l, and 3m having different contents in the thickness direction of the carrier 1 out of the spots (dots) 3, and the contents of the respective detection components. The case where the magnitude of the amount is larger on the other end (Y) side (spot (dot) 3k is the smallest and spot (dot) 3m is the most) is shown.

  By comprising in this way, it can be made to react in the thickness direction of the support | carrier 1, and it can be set as the test piece of a high dynamic range in a small area.

  In 1st invention, you may comprise combining the above-mentioned 1st basic example and 1st modification 1-6 suitably.

  FIG. 8 (a) is an explanatory view schematically showing one basic example (second basic example 1) of the analytical test piece of the second invention, and FIG. 8 (b) is shown in FIG. 8 (a). It is a partial enlarged view of the reaction part (B part) in the test piece for analysis (2nd basic example 1) of 2nd invention shown. Moreover, Fig.9 (a) is explanatory drawing which shows typically the other basic example (2nd basic example 2) of the test piece for analysis of 2nd invention, FIG.9 (b) is FIG.9 (a). FIG. 4 is a partially enlarged view of a reaction part (part D) in the analytical test piece (second basic example 2) of the second invention shown in FIG. As shown in FIGS. 8 (a) and 9 (a), the test specimen 20 for analysis of the second invention comprises a carrier 1 and detection components disposed on and / or inside the surface of the carrier 1. When the sample containing the substance to be analyzed is introduced onto the surface of the carrier 1, it comes into contact with and reacts with the detection component contained in the reaction part 2 and can be detected ( An analytical test piece capable of generating or presenting a detectable substance (signal substance) (signal characteristic), wherein the reaction portion 2 has a plurality of reaction sites 4 and 5 (in FIG. 8B, the reaction site 4a). 4b, 4c, and 4d, and in FIG. 9B, the reaction sites 5a, 5b, 5c, and 5d are shown). From one end (R) to the other end (S), a plurality of reaction sites 4, 5 (see FIG. 8B) In FIG. 9 (b), reaction sites 4a, 4b, 4c, 4d are shown, and in FIG. 9 (b), reaction sites 5a, 5b, 5c, 5d are shown. As shown in FIG. 9 (a), it is configured to increase or decrease, adjacent to each other, or separated from each other, independently and in pieces. Here, “adjacent to each other” means not only the case where the outlines of the reaction sites are arranged in contact with each other, but also the case where the reaction sites are arranged at a slight interval.

  Thus, by providing a difference in the amount per unit area (volume) of the detection component in the reaction portion 2 in the analytical test piece 20, the concentration of the sample containing the substance to be analyzed can be adjusted with one analytical test piece. It can be detected in analog form, and the sample concentration can be digitized. Therefore, a precise value can be output as a test result with a small amount of sample for one analytical test piece. Further, by increasing the area (volume) of the reaction site (by increasing the area where the detection component is the same amount), the determination in the visual inspection can be easily and accurately performed. That is, when making the determination in the visual inspection, it is easy to determine that the width of the region having the same amount is 0.05 mm to 5 mm, more preferably 0.3 mm to 2 mm. When inspecting with an inspection apparatus such as a laser beam, an appropriate size (width) may be set in accordance with the inspection spot diameter of the inspection apparatus. In the case of the second basic example 2, since the content of the detection component in the reaction portion 2 is configured to be separated from each other and independently change (increase or decrease) in a fragmentary manner, an inspection of a laser beam or the like is performed. When automatically inspecting with the apparatus, the inspection can be easily performed. In addition, the size of the discontinuous portion can be appropriately changed in the analytical test piece and applied to alignment, and automatic inspection can be facilitated.

  In the second invention, the same components as in the first invention can be used as the constituent components such as the carrier 1 and the detection component, and the same configuration can be used. In addition, as a modified example (second modified example) of the second invention below, unless otherwise specified, the detection components in each reaction site are configured to increase or decrease stepwise adjacent to each other ( The case shown in FIG.

  FIG. 10 is an explanatory view schematically showing one modified example (second modified example 1) in which the reaction portion in the analytical test piece (second basic example 1) of the second invention shown in FIG. 8 (a) is changed. FIG. As shown in FIG. 10, in the second modification 1, the detection component in the reaction portion 2 is composed of a plurality of types that are spaced apart from each other and arranged independently (the presence of a test component between each type). The detection component content ratio of the detection component from one end (R (A) and R (B)) to the other end (S (A) and S (B)) of the reaction portion. Each of the plurality of reaction sites corresponding to each of the plurality of types is configured to increase or decrease stepwise adjacent to each other. In FIG. 10, four reaction sites composed of two types (A type and B type) of detection components independent from each other (reaction sites 4e, 4f adjacent to each other and a detection component of type B consisting of the detection component of A type). The reaction sites 4g and 4h) adjacent to each other are used. Further, in FIG. 10, the reaction sites 4e and 4f made of the A type detection component have a higher content ratio in the reaction site 4f on the other end (S (A)) side, and the B type detection component. The reaction sites 4g and 4h consisting of the reaction site 4h on the other end (S (B)) side have a higher content ratio.

  With this configuration, a plurality of inspection items can be accurately inspected with a small amount of sample.

  FIG. 11 is a part (part C) of another modified example (second modified example 2) of the reaction portion 2 in the analytical test piece (second basic example 1) of the second invention shown in FIG. 8 (a). It is a partially expanded view which expands and shows typically. As shown in FIG. 11, in the second modified example 1, the detection components in the reaction portion 2 are arranged as an aggregate of spots (dots) 3 with a predetermined arrangement pattern (spot pitch).

  By configuring in this way, similarly to the first modification 2, the sample can be easily supplied from the periphery to the detection component arranged on the surface or inside of the carrier, and the arranged detection component is reacted efficiently. It becomes possible. Therefore, sufficient detection sensitivity can be obtained with a small amount of detection component. Further, since the reaction can proceed from the entire surface of the dot, the detection time can be shortened.

  FIG. 12 schematically shows another modified example (second modified example 3) of the reaction part (B part) in the analytical test piece (second basic example 1) of the second invention shown in FIG. 8 (a). FIG. As shown in FIG. 12, in the second modification 3, the content of the detection component of the spot (dot) 3 constituting the arrangement pattern in the reaction portion 2 is changed from one end (R) to the other end (R) of the reaction portion 2. Up to S) (see FIG. 8A), the plurality of reaction sites 4 that divide the reaction portion 2 are configured to increase or decrease stepwise adjacent to each other. FIG. 12 shows three reaction sites 4i, 4j, and 4k adjacent to each other with different detection component contents of the spot (dot) 3, and the detection component content in the reaction site 4i is the smallest, and the reaction site The case where the content of the detection component in 4k is the largest is shown.

  With this configuration, it is possible to give a change in sensitivity without changing the dot size. Therefore, it can be set as the test piece for analysis of a high dynamic range in a small area.

  FIG. 13 schematically shows another modified example (second modified example 4) of the reaction portion (B part) in the analytical test piece (second basic example 1) of the second invention shown in FIG. 8 (a). FIG. As shown in FIG. 13, in the second modification 4, the arrangement density of spots (dots) 3 constituting the arrangement pattern in the reaction portion 2 is from one end (R) to the other end (S) of the reaction portion 2. (See FIG. 8 (a)), each of the plurality of reaction sites 4 dividing the reaction portion 2 is configured to increase or decrease stepwise adjacent to each other. FIG. 13 shows three reaction sites 41, 4m, and 4n adjacent to each other with different arrangement densities of the spots (dots) 3, and the arrangement intervals of the spots (dots) 3 in the respective reaction sites 41, 4m, and 4n. The pitches p1, p2, and p3 are denser (p1> p2> p3) on the other end (S) side, but are perpendicular to the direction from one end (R) to the other end (S). The pitch of the arrangement interval may be different in any direction, and the pitch of the arrangement interval may be different in both the direction from one end (R) to the other end (S) and the direction perpendicular thereto.

  FIG. 14 schematically shows another modified example (second modified example 5) of the reaction part (B part) in the analytical test piece (second basic example 1) of the second invention shown in FIG. 8 (a). FIG. As shown in FIG. 14, in the second modification example 5, the arrangement size of the spots (dots) 3 constituting the arrangement pattern in the reaction portion 2 is changed from one end (R) to the other end (S ) (See FIG. 8A), the plurality of reaction sites 4 dividing the reaction portion 2 are configured to increase or decrease stepwise adjacent to each other. FIG. 14 shows two reaction sites 4o and 4p adjacent to each other with different sizes of the spots (dots) 3, and the arrangement of the spots (dots) 3o and 3p at the respective reaction sites 4o and 4p. The size of the reaction site 4o is smaller than that of the reaction site 4p.

  By configuring in this way, similar to the first modified example 4 and the first modified example 5, the same concentration and the same amount of detection component (solution) can be easily given a sensitivity difference, so that the productivity is high. Analytical specimens can be obtained and sensitivity differences can be reliably established within the same specimen. In addition, the arrangement pitch and the dot diameter may be set in a range where they do not contact each other in consideration of the sensitivity of the solution, the wettability between the carrier and the solution, etc., but the dot shape cannot be seen in consideration of visual inspection. It is preferable to reduce the distance between dots (the value obtained by subtracting the dot diameter from the pitch) within the range. Specifically, the distance between dots is preferably 0.5 mm or less, and more preferably 0.1 mm or less.

  FIG. 15A shows another modified example (second modified example 6) of the reaction part (B part) in the analytical test piece (second basic example 1) of the second invention shown in FIG. 8A. Fig. 15 (b) is a partially enlarged view schematically showing, and Fig. 15 (b) is a cross-sectional view in the thickness direction of the carrier along the line CC in Fig. 15 (a). As shown in FIG. 15 (a), in the second modification 6, the content in the thickness direction of the carrier 1 in the detected components of the spots (dots) 3 constituting the arrangement pattern in the reaction portion 2 is the reaction. From one end (R) to the other end (S) of the portion (see FIG. 8 (a)), for each of the plurality of reaction sites 4 that divide the reaction portion 2, it is increased or decreased stepwise adjacent to each other. It is configured. In FIG. 15A, three reaction sites 4q adjacent to each other composed of three types of spots (dots) 3q, 3r, and 3s having different contents of detection components in the thickness direction of the carrier 1 are shown. 4r and 4s are shown, and the magnitude of the content of the detection component in each is larger on the other end (S) side (the reaction site 4q is the smallest and the reaction site 4s is the most) Yes.

  By comprising in this way, it can be made to react in the thickness direction of the support | carrier 1, and it can be set as the test piece of a high dynamic range in a small area.

  In the second invention, the above-described second basic examples 1 and 2 and second modified examples 1 to 6 may be combined as appropriate. In addition, a detection component composed of a plurality of independent detection components is arranged in the thickness direction of the carrier, and a substrate in which the solution does not permeate (selectively does not allow the detection element to pass) is arranged between different detection components. By doing so, it is necessary to drop (immerse) the reagent on both surfaces, but it is possible to easily and reliably carry out different types of inspections in a small area.

  In the present invention (the first invention and the second invention), the reaction portion 2 containing the detection component is disposed on the surface and / or inside of the carrier 1 using the ink jet method. preferable.

  In general, when producing a test specimen for analysis, for example, an impregnation method, a QUILL method, a pin and ring method, or a spring pin method is widely used as a method for forming a reaction portion containing a detection component on a carrier. ing.

  The impregnation method is a method in which a detection component (solution) is immersed in a carrier and then dried so that the detection component (solution) is disposed on and / or inside the carrier, and then the detection component is immobilized. However, in this method, the high precision, high density, and high sensitivity required for the test specimens for analysis in recent years (specifically, a reaction part having a concentration difference or density difference of detection components is formed. There is a problem that it is difficult to cope with the above.

  In the QUILL method, a detection component (solution) is stored in a recess formed at the tip of a pin, and the detection component (solution) in the recess is moved onto the support by bringing the pin tip into contact with the support to form a spot (reaction portion). It is a method to do. However, with this method, the pin tip is deformed or damaged due to contact with the carrier, and the cleaning of the detection component (solution) accumulated in the recess is incomplete and cross contamination. There are problems such as being easy to occur.

  In the pin-and-ring method, the detection component (solution) in the microplate is reserved by the ring, and then the detection component (solution) passes through the reserved ring, and the detection component (solution) in the ring at the pin tip And spot (reaction part) is formed on the carrier. However, in this method, the detection components (solutions) that can be reserved at one time depend on the number of rings, and since the number of detection components (solutions) has been about several in the past, detection components (solutions) of several thousand to tens of thousands In order to form a small spot (reaction portion), a washing and drying process of several hundred to several thousand times is also necessary, and therefore, it is difficult to say that productivity is high. Further, there is a problem that the detection component (solution) spreads on the carrier (solution amount) is not reproducible.

  The spring pin method is a method in which the detection component (solution) attached to the pin tip is moved onto the carrier by pressing the pin tip against the carrier to form a minute spot (reaction part). The pin structure softens damage to the pin and carrier and blows out the detection component (solution). However, this method is basically inferior in productivity because only one spotting can be performed with one reservation. Furthermore, these conventional methods for forming microspots (reaction parts) all carry the detection component (solution) on the carrier while being exposed to the atmosphere, so that the detection component (solution) dries in the middle of transportation and spotting occurs. There is a problem that the use efficiency of a very expensive detection component (solution) is poor due to the inconvenience that it becomes impossible.

  On the other hand, a solution containing a detection component is supplied to the surface of the carrier 1 in a non-contact state by disposing the reaction portion 2 on and / or inside the surface of the carrier 1 using an ink jet method. Therefore, the test specimen for analysis can be obtained at high speed and efficiently without damaging the carrier 1.

  In addition, by using the ink jet method, it is possible to increase the accuracy, density, and sensitivity of the position of the reaction part, for example, a spot (dot) in a predetermined arrangement pattern, and the amount of liquid input for each spot. (Spot amount) can be made accurate. Therefore, it is possible to accurately match the predetermined sensitivity in the analytical test piece, improve the reliability of the analysis, maintain the uniformity of quality even if the analytical test piece is enlarged, and improve production efficiency. Can be achieved. In addition, it is possible to obtain a highly sensitive analytical test piece that can be detected even with a small amount of sample by increasing the density. Furthermore, since the amount of the input liquid can be made accurate, the measurement error of the obtained analytical test piece can be reduced, and the measurement fluctuation can be suppressed.

  As a droplet discharge device used in the ink jet method, for example, a flow path base in which a flow path is formed, and an actuator unit that is assembled to the flow path base and has a function of changing the volume in the cavity as a pressurizing chamber; , One provided with a nozzle base adhered to the lower surface of the flow path base and having a discharge nozzle formed thereon, and a liquid injection part provided on the upper surface of the rear part of the flow path base (hereinafter referred to as “discharge unit”) Can be mentioned). Specifically, one discharge unit or a plurality of discharge units described in Japanese Patent Application Laid-Open No. 2003-75305 can be suitably used.

  When using a plurality of discharge units, a test piece can be produced efficiently by forming a plurality of dots simultaneously. In addition, it is preferable to make the difference in the discharge amount by making the size of each discharge nozzle different from each other, because it is easy to make a difference in the detected component amount. In addition, the discharge amount may be varied by changing the amount or speed of changing the volume in the cavity in each discharge unit, and the difference in height between the liquid level of the liquid injection part and the discharge nozzle may be set in each discharge unit. It is also preferable to make a difference in the discharge amount between the discharge devices by making a difference between them.

  Further, as a system for controlling such a droplet discharge device, for example, a system described in JP-A-2003-98183 can be suitably used. In addition to the ink jet method, a screen printing method in which the mesh aperture ratio is changed in consideration of the viscosity of the solution can be used as appropriate.

  In the present invention, a support for supporting the carrier may be further disposed on the surface (back surface) side opposite to the surface of the carrier. By doing in this way, handling is easy and the arrangement | sequence (alignment) at the time of forming a spot (dot) can be facilitated. In addition, as a support body, a metal, ceramics, glass, resin etc. can be mentioned as a material, for example.

  Further, a confirmation reaction part for confirming in advance that the detection component reacts normally may be provided outside the reaction part on the carrier. In addition, in order to facilitate the determination in the visual inspection, from one end (X) or (R) of the reaction portion to the other end (Y) or (S), for each of a plurality of reaction sites 4 that divide the reaction portion 2, A marking may be placed in each reaction site 4 that is configured to increase or decrease continuously, stepwise or discontinuously and in pieces. As such marking, for example, no detection component can be arranged at the location.

  EXAMPLES Hereinafter, the present invention will be described more specifically with reference to examples, but the present invention is not limited to these examples. In this example, an analytical test piece was prepared using the inkjet method, and it was examined whether or not analog detection could be performed with the same amount of test solution.

Example 1
As a carrier, a material having a hydrophilic cellulose mixed ester, a size of 5 mm × 5 mm, a pore diameter of 0.8 μm, and a thickness of 0.16 mm was used, and a solution containing a detection component placed on this carrier was 100 units / ml glucose dehydrogenase, 20 mM β-NAD (nicotinamide adenine dinucleotide), 20 units / ml diaphorase, 20 mM MTT (3- (4,5-dimethylthiazol-2-yl) -2,5 -Diphenyl-2H-tetrazolium bromide), and further, as a method of forming a reaction portion, a discharge unit (inkjet method) described in Japanese Patent Application Laid-Open No. 2003-75305 is used. Was made. This sample for evaluation has 8 reaction sites in 3 mm × 3 mm within a size of 5 mm × 5 mm (having an analog difference in sensitivity in 8 steps), and the components per unit area of each reaction site in the reaction part the amount (liquid amount of the solution) was assumed that the 8 steps from 1 nL / mm ² to 8 nL / mm ² was continuously increased. In addition, as a sample containing the substance to be analyzed, a solution with glucose content as a parameter (glucose content 10 mg / dl, 20 mg / dl, 50 mg / dl) was prepared, and the sample carrier for evaluation was prepared. It applied to the surface and the sensitivity characteristic of the sample for evaluation was evaluated by visual inspection. The results are shown in Table 1. As shown in Table 1, blue color development in the state shown in Table 1 was confirmed for each reaction site (component amount (solution amount)) in the reaction portion of the sample for evaluation. In Table 1, symbol x indicates that no blue color is visible, symbol Δ indicates that it is somewhat visible, and symbol O indicates that it is visible.

  As can be seen from Table 1, one analysis test piece (one analysis configured such that the content ratio of the detection component in each reaction site increases continuously from one end to the other end of the reaction portion. The test specimen) confirmed that the amount of glucose was detected in an analog fashion.

(Example 2)
In Example 1, tris containing 10 units / ml cholesterol dehydrogenase, 5 mM NAD (nicotinamide adenine dinucleotide), 20 units / ml diaphorase, 5 mM NTB as a solution containing a detection component to be arranged on a carrier. A sample dissolved in a buffer solution (pH 8.0) was used, and as a sample containing a substance to be analyzed, the cholesterol content was used as a parameter (the cholesterol content was 20 mg / dl, 40 mg / dl, 100 mg / dl). A sample for evaluation was prepared in the same manner as in Example 1 except that the solution was used, and the sensitivity characteristics of the sample for evaluation were evaluated by visual inspection in the same manner as in Example 1. The results are shown in Table 2. As shown in Table 2 (the meanings of the symbols in Table 2 are the same as those in Table 1), each reaction site (component amount (liquid amount of the solution)) is divided into Table 2 A reddish purple color was confirmed in the state shown in FIG.

  As can be seen from Table 2, one analytical test strip (one analysis configured such that the content ratio of the detection component in each reaction site increases continuously from one end of the reaction portion to the other end). The test specimen) confirmed that the amount of cholesterol was detected in an analog fashion. Moreover, in Example 1 and Example 2, although the test piece for analysis (sample for evaluation) was produced by the inkjet method, a screen printing method, a liquid impregnation method, or the like may be appropriately selected and used.

  The test specimen for analysis of the present invention examines and analyzes the properties of a sample containing a substance to be analyzed (for example, body fluids of humans and animals, particularly urine, blood, etc.) in the fields of research, drug discovery, diagnosis, medical treatment, etc. It is effectively used for manufacturing inspection chips and the like.

Claims (12)

Comprising a carrier and a reaction portion containing a detection component disposed on and / or in the surface of the carrier, and when a sample containing an analyte is introduced into the surface of the carrier, contacting said detection components contained in the reaction section, react to a analytical test strip capable of generating or detectable characteristics of presentation of detectable objects quality,
The content ratio of the detection component in the reaction portion is configured to continuously increase or decrease from one end to the other end of the reaction portion,
The detection component in the reaction portion, it becomes disposed in a predetermined arrangement pattern as a collection of spots, the arrangement density of the spots constituting the arrangement pattern, in from one end of the reaction section to the other end An analytical test strip configured to continuously increase or decrease.   The detection component in the reaction part is composed of a plurality of types that are spaced apart from each other and are arranged independently, and the content of the detection component is from one end to the other end of the reaction part for each type of the detection component. The analytical test strip according to claim 1, wherein the analytical test strip is configured to continuously increase or decrease. The content of the detected components of spots constituting the arrangement pattern, the in from one end of the reaction portion to the other end, the analysis of continuously increasing or configured to decrease composed according to claim 1 or 2 Test piece. The size of the spots constituting the arrangement pattern, in from one end of the reaction portion to the other end, for analysis according to any one of the continuously become configured to increase or decrease according to claim 1 to 3 Test pieces. In the detection component of spots constituting the arrangement pattern, the content in the thickness direction of the carrier, in from one end of the reaction section to the other, consisting configured to increase or decrease continuously claims Item 5. The test specimen for analysis according to any one of Items 1 to 4. Comprising a carrier and a reaction portion containing a detection component disposed on and / or in the surface of the carrier, and when a sample containing an analyte is introduced into the surface of the carrier, contacting said detection components contained in the reaction section, react to a analytical test strip capable of generating or detectable characteristics of presentation of detectable objects quality,
The reaction portion is configured to be divided into a plurality of reaction sites, and the content ratio of the detection component in the reaction portion is adjacent to each other for each of the plurality of reaction sites from one end to the other end of the reaction portion. It is configured to increase or decrease in stages or apart from each other, independently of each other, and
The detection component in the reaction portion, it becomes disposed in a predetermined arrangement pattern as a collection of spots, the arrangement density of the spots constituting the arrangement pattern, in from one end of the reaction section to the other end An analytical test strip configured to increase or decrease continuously, stepwise, or discontinuously and fragmentally for each of the plurality of reaction sites that divide the reaction portion.   The detection component in the reaction part is composed of a plurality of types that are spaced apart from each other and arranged independently, and the content ratio of the detection component is a plurality of the detection components from one end to the other end of the reaction part. 7. The analytical device according to claim 6, wherein each of the plurality of reaction sites corresponding to each type is configured to increase or decrease stepwise adjacent to each other, or separated from each other and independently fragmented. Test pieces. The content of the detected components of spots constituting the arrangement pattern, in from one end of the reaction portion to the other end, for each of the plurality of the reaction sites that divide the reaction portion, and stepwise continuous or non The analytical test strip according to claim 6 or 7, wherein the analytical test strip is configured to increase or decrease continuously and fragmentarily. The size of the arrangement of the spots constituting the arrangement pattern, in from one end of the reaction portion to the other end, for each of the plurality of the reaction sites that divide the reaction portion, stepwise or discontinuously in succession The analytical test strip according to any one of claims 6 to 8, wherein the analytical test strip is configured to increase or decrease in a fractional manner. In the detection component of spots constituting the arrangement pattern, the content in the thickness direction of the carrier, in from one end of the reaction portion to the other end, for each of the plurality of the reaction sites that divide the reaction portion, The analytical test strip according to any one of claims 6 to 9, wherein the analytical test strip is configured to increase or decrease continuously, stepwise, or discontinuously and fragmentarily.   The analytical test piece according to claim 1, wherein the carrier is a fibrous body or a porous body.   The analytical test piece according to any one of claims 1 to 11, wherein the reaction portion containing the detection component is arranged on the surface and / or inside of the carrier using an inkjet method.

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